TLRs make up a family of pattern recognition receptors that play important roles in the innate immune response. Broad classes of pathogens (e.g. viruses, bacteria, and fungi) constitutively express a set of mutation-resistant molecules called pathogen-associated molecular patterns (PAMPs). These microbial molecular markers may be composed of proteins, carbohydrates, lipids, nucleic acids and/or combinations thereof. Individual TLRs recognize distinct pathogen-associated PAMPs, initiating signaling cascades that promote the immune response. Structurally, TLRs are type I transmembrane receptors that possess varying numbers of extracellular N-terminal leucine-rich repeat (LRR) motifs, followed by a cysteine-rich region, a TM domain, and an intracellular Toll/IL-1 R (TIR) motif. The TIR motif is common to the larger IL-1 R/TLR superfamily.
Discontinued Product
DTLR20 has been discontinued.
View all TLR2 products.
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Cell Lysates (10 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Saliva (34 µL), Urine (100 µL), Human Milk (10 µL)
Sensitivity
0.109 ng/mL
Assay Range
0.156-10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human TLR2 Quantikine ELISA Kit
The Quantikine Human TLR2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human TLR2 in cell culture supernates, cell lysates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human TLR2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TLR2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human TLR2.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human TLR2.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, Human Milk, Saliva, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 1.32 | 2.72 | 5.47 | 1.40 | 2.82 | 5.73 |
| Standard Deviation | 0.032 | 0.055 | 0.109 | 0.105 | 0.225 | 0.427 |
| CV% | 2.4 | 2.0 | 2.0 | 7.5 | 8.0 | 7.5 |
EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 1.43 | 3.01 | 5.82 | 1.48 | 3.12 | 6.20 |
| Standard Deviation | 0.051 | 0.065 | 0.155 | 0.128 | 0.310 | 0.460 |
| CV% | 3.6 | 2.2 | 2.7 | 8.6 | 9.9 | 7.4 |
Recovery for Human TLR2 Quantikine ELISA Kit
The recovery of human TLR2 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 96 | 92-98 |
| EDTA Plasma (n=4) | 98 | 86-107 |
| Heparin Plasma (n=4) | 98 | 88-105 |
| Serum (n=4) | 99 | 84-111 |
| Urine (n=4) | 98 | 91-108 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human TLR2 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human TLR2 Quantikine ELISA Kit
Human TLR2 ELISA Serum/Plasma Standard Curve
Human TLR2 ELISA Cell Culture Supernate/Cell Lysate/ Saliva/Urine/Human Milk Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TLR2
Long Name
Toll-like Receptor 2
Alternate Names
CD282
Gene Symbol
TLR2
Additional TLR2 Products
Product Documents for Human TLR2 Quantikine ELISA Kit
Product Specific Notices for Human TLR2 Quantikine ELISA Kit
For research use only
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Protocols
View specific protocols for Human TLR2 Quantikine ELISA Kit (DTLR20):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Human TLR2 Quantikine ELISA Kit
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A: Assay Diluent RD1-75 contains high concentrations of additives and typically has a very cloudy, brown appearance. It is recommended to mix this diluent well before and during use.
